Association of double-stranded RNA with whitefly-mediated silvering in squash

A disorder of unknown aetiology appearing as silvering on squash (Cucurbita pepo) leaves has become prevalent in south Florida. Typical symptoms, observed after 2 weeks of exposure to adults of Bemisia tabaci , the sweet potato whitefly (SPWF) include vein clearing followed by silvering of whole leaves. Fruits from affected plants are blanched or streaked longitudinally. The silverleaf syndrome was not mechanically transmissible when crude extracts of symptomatic leaves were used as inoculum. Two sizes of double-stranded RNA (dsRNA) of c. 4.6 and 4.2 kilobase pairs (kbp) were consistently observed in leaf extracts of caged, whitefly-infested, symptomatic plants. In contrast, caged plants not exposed to whiteflies showed no symptoms and contained no detectable dsRN A. In addition, squash plants exposed to SPWF colonies from California neither developed silverleaf symptoms nor contained any dsRNA. Double-stranded RNA could also be readily detected in whitefly-infested yellow summer squash and zucchini from the field. However, asymptomatic eggplant and watermelon infested with SPWF from nearby fields did not contain detectable dsRNA. Dot-spot hybridization assays using an RNA-specific probe detected homologous sequences in both adults and nymphs of SPWF that induced silverleaf symptoms but not in adults and nymphs of SPWF that could not induce silverleaf symptoms. These results, along with the non-endogenous nature of the detected dsRNA suggest that the dsRNA is or is associated with the causal agent of whitefly-mediated leaf silvering in squash.